A granulocyte-macrophage colony stimulating factor (GM-CSF) has an action for stimulating the proliferation and differentiation of granulocyte progenitor and is a glycoprotein having a molecular weight of 22,000, which produces neutrophils, acidocytes and monocytes within an organism.
As the diseases associated with a blood state of low GM-CSF level, there are mentioned neutropenia, aplastic anemia, osteomyelodysplasia syndrome, or the like, and GM-CSF is in use for treatment of these diseases (e.g. Hideaki Mizoguchi, GM-CSF, Cytokine Therapy: Approach from Basis and Pathema, Nankodo Co., Ltd., Tokyo, p. 39-45, 1993, ISBN: 4524233628). Also, it is shown that two cytokines of granulocyte colony stimulating factor (G-CSF) and GM-CSF stimulate the proliferation of granulocyte progenitor fraction and shorten the duration of decrease of neutrophil, and they are in use as first-line drugs for leucopenia and granulopenia (e.g. Merck Manual 16th Edition, p. 1240, 1994).
Tumor necrosis factor (TNF) was reported for the first time as a factor which kills tumor cells in vitro. Thereafter, lymphotoxin-α (LT-α) and lymphotoxin-β (LT-β) were identified as factors having similar bioactivities. TNF is a general term of molecules having a tumor cell-killing effect, and there are known three kinds, i.e. TNF-α, TNF-β (LT-α) and LT-β. TNF bonds with receptors present in almost all cells within an organism and thereby exhibits extensive actions (e.g. Tsutomu Takeuchi, Separate Volume: Progress of Medical Science, Immunodeficiency, Ishiyakushuppan Co., Ltd., Anti-TNF α Therapy, p. 538-542, 2002).
TNF is a multifunctional cytokine which exhibits different activities to various kinds of cells in vitro tests. As the diseases caused by an increase in TNF-α, there are known rheumatoid arthritis (RA), Crohn's disease and inflammatory bowel disease (IBD). For example, it is reported that, in patients of rheumatoid arthritis, TNF-α reaches 15.0±9.2 pg/ml (e.g. Masahiro Yamamura, Clinical Analysis and Treatment of Anemia in Patients of Rheumatoid Arthritis, Report for 6th, Incentive Award of the Society of Certifying Specialist in Internal Medicine, 1998) and, in Crohn's disease, the average TNF-α of four patients in inactive period is 11.98 pg/ml and the average TNF-α of four patients in active period is 404.76 pg/ml (e.g. Naoki Watanabe, Development of Highly Sensitive Assay for Detection of Intravital Trace Materials and Its Clinical Application, Lab. Clin. Pract., 20(2), p. 110-114, 2002).
RA is a cryptogenic chronic inflammatory disease which appears mainly at synovial membrane of joint. It was made clear that, in articular cavity, inflammatory cytokines such as TNF-α, IL-1, IL-6 and the like are produced in excess and participate in formation of joint lesion such as lymphocytic infiltration, synovial hyperplasia and destruction of cartilage tissue by osteoclast. Arthritis was caused in a model animal in which was high expression of TNF-α and, when the TNF-α was neutralized with an anti-TNF-α monoclonal antibody, striking improvement of the arthritis was seen. In association therewith, IL-1 and IL-6 concentrations decreased. Based on this, it has considered that TNF-α has a close connection with excessive production of the above inflammatory cytokines and plays a central role in formation of RA lesion at an upstream of cytokine cascade (e.g. Tsutomu Takeuchi, Appendix: Progress of Medical Science, Immunodeficiency, Anti-TNF α Therapy, Ishiyakushuppan Co., Ltd., p. 538-542, 2002). The anti-TNF-α monoclonal antibody was approved as a remedy for rheumatoid already in 1999 in U.S.A.
Crohn's disease is a disease in which inflammation of unknown etiology persists mainly in the small intestine and the large intestine and induces the beginning of ulcer, the sequential expression of stenosis, abscess and fistula in the intestinal tract. Crohn's disease is often difficult to differentiate from ulcerative colitis and, together with ulcerative colitis, is generally called as Inflammatory Bowel Disease (IBD). In treatment of Crohn's disease, it is important to effectively control the persisting inflammation of intestinal tract. Substances associated with inflammation include several tens of kinds and, of them, TNF-α has been clarified to have a central role. A large amount of TNF-α is produced and present when the inflammatory intestinal duct of a patient of Crohn's disease is examined. This large amount of TNF-α cause further inflammation; therefore, in order to break out of the vicious cycle of inflammation, counteractive to the effect of TNF-α have been developed. Also, a thalidomide therapy has been applied to refractory Crohn's disease and the effectiveness of thalidomide to some cases was proven (e.g. Ehrenpreis E D, Thalidomide therapy for patients with refractory Crohn's disease: An open-label trial, Gastroenterology, 117, p. 1271-1277, 1999). Also, a chimeric monoclonal anti-TNF antibody (Infliximab, manufactured by Centocor, Inc., U.S.A.) was proven as a first remedy by biotechnology in the field of gastroenterological disease, in 1999 in U.S.A. It is reported that the treatment using the remedy suppresses the inflammation of Crohn's ileocolitis significantly (e.g. Baert FL. Tumor necrosis factor α-antibody (Infliximab) therapy profoundly down-regulates the inflammation in Crohn's ileocolitis. Gastroenterology, 116, p. 22-28, 1999). In Japan, the above antibody was approved as a remedy for Crohn's disease in January, 2002.
Meanwhile, as to Cucurbita moschata, Carthamus tinctorius, Plantago asiatica and Lonicera japonica, respective efficacies are reported as follows. For example, it is disclosed that addition, to a feed, of at least one kind of Cucurbita moschata, Plantago asiatica and Lonicera japonica (particularly, a crude drug comprising these three crude drugs) can prevent, in particular, natural infection of parasites, bacteria and viruses and can achieve higher biophylaxis ability and improved meat and egg qualities. Further, it has been disclosed that a feed comprising four kinds of crude drugs, i.e. Cucurbita moschata, Plantago asiatica, Lonicera japonica and Carthamus tinctorius improves the health conditions, survival rates, quality of egg, and has anti-leucocytozoonosis effect in layers, and anti-New-Castle-disease effect and effects of the decreased numbers of enteric Coccidium and Staphylococcus in the intestine of quails (e.g. U.S. Pat. No. 5,882,672).
A method for producing an interferon inducer from the plants of the genus Cucurbitaceae such as pumpkin has been disclosed (e.g. U.S. Pat. No. 4,421,746). The antiviral activity and anti-tumor activity of interferon inducers extracted from the flowers of Carthamus tinctorius has been disclosed (e.g. U.S. Pat. No. 4,456,597). It has also been disclosed that interferon inducers may be extracted from the flowers of Lonicera japonica, seeds of Plantago asiatica, and the like, and that the extracted interferon inducers are useful for prevention and curative treatments of viral infections in humans and animals (e.g. U.S. Pat. No. 4,469,685). A macrophage activator comprising two crude drugs of Cucurbita moschata and Carthamus tinctorius has also been disclosed (e.g. Japanese Patent Laid-open No. 116498/1999). A neutrophil activator comprising four crude drugs of Cucurbita moschata, Carthamus tinctorius, Plantago asiatica and Lonicera japonica has also been disclosed (e.g. Japanese Patent Laid-open No. 281584/2000).
However, while these documents disclose the interferon inducing effects, macrophage activating effects, neutrophil activating effects, the inhibitory effects of IgE anti-body production, and the like of the crude drugs used as active components in the present invention, none of the references disclose or suggest the increasing action for GM-CSF concentration in blood or on the proliferating action for granulocyte or macrophage.
The aim of the present invention is to provide a GM-CSF increasing agent comprising, as active components, crude drugs, specifically, Cucurbita moschata, Carthamus tinctorius, Plantago asiatica and Lonicera japonica. 